GSM7658516: uninfected_control_2_S5; Mus musculus; RNA-Seq
After euthanasia, the lungs were extracted, washed with PBS to remove excess blood. Ca. 60 mg of tissue was added to 1 mL of Trizol and maintained on ice. The samples were then homogenized using zirconia beads and a Precellys Beadbeater and the nucleic acids were isolated after addition of chloroform and precipitation of the resulting acqeous layer with an equal volume of 70% ethanol. Thhereafter the samples were removed from the BSL-3 laboratory and RNA extracted using the Qiagen RNeasy Mini Kit with on-column DNase treatment Libraries were prepared from ribodepleted RNA isolated from lung tissue taken from Mtb-infected mice and sequenced using the NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (New England Biolabs) as per the manufacturer's instructions. A mean of 51.9 million 60 base-pairs paired-end reads were obtained among the 9 samples (39.2 – 69.3 million reads).